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Updated: Sep 9, 2025

"Phagosome Closure Assay" to Visualize Phagosome Formation in Three Dimensions Using Total Internal Reflection Fluorescent Microscopy TIRFM
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Physical confinement and phagocytic uptake induce persistent cell migration.

Summer G Paulson1,2, Sophia Liu1,2, Jeremy D Rotty1

  • 1Uniformed Services University of the Health Sciences, Department of Biochemistry, Bethesda, MD, USA, 20814.

Biology Open
|September 1, 2025
PubMed
Summary
This summary is machine-generated.

Physical confinement significantly enhances phagocytosis in BV2 microglia-like cells, acting as a potent driver. This effect involves the Arp2/3 complex and myosin II, influencing cell migration and immune responses.

Keywords:
ActinArp2/3 complexConfined motilityFibronectinMicrogliaMyosinPhagocytosis

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Area of Science:

  • Cell Biology
  • Immunology
  • Biophysics

Background:

  • Phagocytosis is typically studied in unconfined in vitro settings.
  • The influence of physical confinement on phagocytosis is not well understood.
  • Microglia play a critical role in the innate immune response.

Purpose of the Study:

  • To investigate the impact of physical confinement on IgG-mediated phagocytosis in BV2 microglia-like cells.
  • To elucidate the cytoskeletal mechanisms underlying confinement-modulated phagocytosis and cell migration.
  • To explore the phenomenon of 'phagocytic priming' and its dependence on confinement and cell adhesion.

Main Methods:

  • Utilized BV2 microglia-like cells in both confined and unconfined in vitro assays.
  • Investigated IgG-mediated phagocytosis using fluorescent beads.
  • Examined the roles of Arp2/3 complex, myosin II, and integrin-dependent adhesion using pharmacological inhibitors and genetic disruption.
  • Assessed cell migration and 'phagocytic priming' using live-cell imaging.

Main Results:

  • Physical confinement significantly increased the fraction of phagocytic cells compared to unconfined conditions.
  • Confinement partially rescued phagocytic uptake upon myosin II disruption and conferred partial resistance to cytochalasin D.
  • Bead uptake stimulated persistent cell migration, termed 'phagocytic priming', which required integrin-dependent adhesion.
  • Cytoskeletal requirements for phagocytic priming differed between confined (Myosin II, Arp2/3 complex required) and unconfined settings.

Conclusions:

  • Physical confinement is a potent enhancer of phagocytosis in microglia.
  • Confinement alters the cytoskeletal dynamics of phagocytosis and phagocytic priming.
  • Phagocytic priming, modulated by confinement, may be a key innate immune mechanism for wound surveillance.